Question (9589) | Jurkat nuclear extract lysate (ab14844)

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Question

I'd like to use this nuclear lysate for my PPAR-gamma, delta Western-blot as a positive control. Do I have to boil it up or use any kind of pre-treatment with this lysate or it is ready to load. Typically how many microgramms do you suggest to load for a big gel to be able to see the proteins, mentioned above?

Answer

Thank you for your enquiry and your interest in our products. Ab14844 is Jurkat nuclear lysate and you need to dilute it in a standard Western blot sample buffer. It may be worth boiling the sample for 1 or 2 minutes before loading. Usually for mini SDS gel, we suggest loading 20-30 ug protein per lane, whilst for big gel you may need to load 50 ug. It depends on the exact size of the gel and the comb you use. We would suggest taking a look at the manufacturer's instructions. We hope this will help.

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